The objective of Phase I studies proposed herein is to develop reagents and methods to rapidly and unambiguously screen the human genome to validate genes as drug discovery targets. In phase II studies, the methods will be used to discover and validate targets to pursue for treatment of pancreatic cancer, although the technology is applicable for other indications. Gene knockouts have been considered the most unambiguous means for inactivating genes. The ability to screen libraries of cells containing inactivated genes to demonstrate functions of genes has proven to be highly successful, but has typically been reserved for non-mammalian cells. The innovative technology described in the proposed research allows for gene knockouts and screening capabilities to enable gene evaluations to now be performed in high throughput with mammalian cell lines. Specific aims proposed are 1) generation of a 2,000 gene library for gene knockouts, 2) creation of 10 gene knockout vectors, 3) inactivation of 10 genes in 2 human pancreatic cell lines, and 4) evaluation of phenotypic effects. Creation of large libraries of genes and screening these libraries in well-defined assays will enable discovery of genes, their biochemical functions, their roles in signaling networks relevant to human diseases, and their therapeutic utilities. PROPOSED COMMERCIAL APPLICATION: PanGenex plans to capitalize on its innovative and proprietary technology both in a service capacity (with potential short-term and long-term revenue projections) and to provide for sustained drug discovery programs both within PanGenex and with corporate partners.